Isolation and characterization of ornitho‐kininogen

Makoto Kimura, Tatsuya SUEYOSHI, Katsumi TAKADA, Kosaku TANAKA, Takashi MORITA, Sadaaki IWANAGA

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Abstract

Ornitho‐kininogen was purified from chicken blood plasma by a two‐stage method using chromatography on columns of S‐alkylated papain‐Cellulofine and DEAE‐5PW. The yield was 1.7 mg from 44 ml plasma. The isolated preparation gave a single band on sodium dodecyl sulfate/polyacrylamide gel electrophoresis (SDS‐PAGE) with or without 2‐mercaptoethanol and on disc/polyacrylamide gel electrophoresis. The relative molecular mass, Mr, of ornitho‐kininogen was estimated as 74000 on SDS‐PAGE using the Ferguson plot method. Ornitho‐kininogen was found to have the similar properties to those of mammalian high‐Mr kininogen, in terms of the amino acid composition, molecular mass, and susceptibility to plasma kallikrein. No kininogen corresponding to mammalian low‐Mr kininogen and rat T‐kininogen could be detected in chicken plasma. In fact, ornitho‐kininogen was degraded rapidly by bovine plasma kallikrein, liberating a kinin. This kinin was isolated from the digest by reversed‐phase HPLC. The primary structure of the isolated kinin was determined as Arg1‐Pro2‐Pro3‐Gly4‐Phe5‐Thr6‐Pro7‐Leu8‐Arg9. The sequence of this peptide, named ornitho‐kinin, was similar to that of bradykinin except for the substitution of Thr6 and Leu8 for Ser6 and Phe8. The isolated ornitho‐kinin induced a contraction of chicken smooth muscle and had a strong hypotensive effect in the chicken. However, it did not contract the isolated rat uterus. It is suggested that this specificity difference is due to the replacement of Phe8 by Leu8. The sequence of residues 1–30 of ornitho‐kininogen exhibited 43% identity with that of bovine kininogen.

Original languageEnglish
Pages (from-to)493-499
Number of pages7
JournalEuropean Journal of Biochemistry
Volume168
Issue number3
DOIs
Publication statusPublished - Jan 1 1987

All Science Journal Classification (ASJC) codes

  • Biochemistry

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    Kimura, M., SUEYOSHI, T., TAKADA, K., TANAKA, K., MORITA, T., & IWANAGA, S. (1987). Isolation and characterization of ornitho‐kininogen. European Journal of Biochemistry, 168(3), 493-499. https://doi.org/10.1111/j.1432-1033.1987.tb13444.x